Skip to main content
SpringerLink
Log in

Polyunsaturated fatty acids support epithelial barrier integrity and reduce IL-4 mediated permeability in vitro

  • ORIGINAL CONTRIBUTION
  • Published:
European Journal of Nutrition Aims and scope Submit manuscript

Abstract

Background

The intestinal mucosa functions as a barrier against harmful dietary and microbial antigens. An intact gut barrier forms a prerequisite for protection against infection and allergy. Both allergic and inflammatory mediators (e.g. IL-4, IFN-γ) are known to compromise the epithelial barrier integrity by enhancing permeability. Breast milk provides protection against infection and allergy and contains polyunsaturated fatty acids (PUFA).

Aim of the study

Although PUFA are commonly used in infant formulas their effect on intestinal barrier is still poorly understood. Therefore the effects of distinct PUFA (n-6: LA, GLA, DGLA, AA; n-3: ALA, EPA, DHA) and a fat blend with PUFA composition similar to that of the human breast milk fat fraction, on barrier integrity were investigated.

Methods

Human intestinal epithelial cells (T84) were pre-incubated with individual PUFA or a lipase treated fat blend, with or without subsequent IL-4 exposure. Barrier integrity was evaluated by measuring transepithelial resistance and permeability. Membrane phospholipid composition was determined by capillary gas chromatography.

Results

DGLA, AA, EPA, DHA and to a lesser extend GLA enhanced basal TER and strongly reduced IL-4 mediated permeability, while LA and ALA were ineffective. Furthermore, the lipase treated fat blend effectively supported barrier function. PUFA were incorporated in the membrane phospholipid fraction of T84 cells.

Conclusions

Long chain PUFA DGLA, AA, EPA and DHA were particularly effective in supporting barrier integrity by improving resistance and reducing IL-4 mediated permeability. Fat blends that release specific PUFA upon digestion in the gastrointestinal tract may support natural resistance.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

FD4:

4 kDa FITC-dextran

IL-4:

Interleukin-4

(LC-) PUFA:

(Long chain-)polyunsaturated fatty acids

TER:

Transepithelial resistance

References

  1. Alessandri JM, Guesnet P, Arfi TS, Durand G (1991) Changes in fatty acid composition during cell differentiation in the small intestine of suckling piglets. Biochim Biophys Acta 1086: 340–348

    CAS  Google Scholar 

  2. Alessandri J-M, Joannic J-L, Durand GA (1993) Polyunsaturated fatty acids as differentiation markers of jejunal epithelial cells: a modeling approach. J Nutr Biochem 4:97–104

    Article  CAS  Google Scholar 

  3. Barton RG, Cerra FB, Wells CL (1992) Effect of a diet deficient in essential fatty acids on the translocation of intestinal bacteria. JPEN J Parenter Enteral Nutr 16:122–128

    Article  CAS  Google Scholar 

  4. Benard A, Desreumeaux P, Huglo D, Hoorelbeke A, Tonnel AB, Wallaert B (1996) Increased intestinal permeability in bronchial asthma. J Allergy Clin Immunol 97:1173–1178

    Article  CAS  Google Scholar 

  5. Berin MC, Yang PC, Ciok L, Waserman S, Perdue MH (1999) Role for IL-4 in macromolecular transport across human intestinal epithelium. Am J Physiol 276:C1046–C1052

    CAS  Google Scholar 

  6. Beyer K, Castro R, Birnbaum A, Benkov K, Pittman N, Sampson HA (2002) Human milk-specific mucosal lymphocytes of the gastrointestinal tract display a TH2 cytokine profile. J Allergy Clin Immunol 109:707–713

    Article  CAS  Google Scholar 

  7. Blight EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Med Sci 37:911–917

    Google Scholar 

  8. Caplan MS, Russell T, Xiao Y, Amer M, Kaup S, Jilling T (2001) Effect of polyunsaturated fatty acid (PUFA) supplementation on intestinal inflammation and necrotizing enterocolitis (NEC) in a neonatal rat model. Pediatr Res 49:647–652

    Article  CAS  Google Scholar 

  9. Catassi C, Bonucci A, Coppa GV, Carlucci A, Giorgi PL (1995) Intestinal permeability changes during the first month: effect of natural versus artificial feeding. J Pediatr Gastroenterol Nutr 21:383–386

    Article  CAS  Google Scholar 

  10. Chawla A, Karl PI, Fisher SE (1995) Effect of N-3 polyunsaturated fatty acid supplemented diet on neutrophil-mediated ileal permeability and neutrophil function in the rat. J Am Coll Nutr 14:258–263

    CAS  Google Scholar 

  11. Colgan SP, Resnick MB, Parkos CA, Delp-Archer C, McGuirk D, Bacarra AE, Weller PF, Madara JL (1994) IL-4 directly modulates function of a model human intestinal epithelium. J Immunol 153:2122–2129

    CAS  Google Scholar 

  12. DeMeo MT, Mutlu EA, Keshavarzian A, Tobin MC (2002) Intestinal permeation and gastrointestinal disease. J Clin Gastroenterol 34:385–396

    Article  Google Scholar 

  13. Dooper MM, Wassink L, M’Rabet L, Graus YM (2002) The modulatory effects of prostaglandin-E on cytokine production by human peripheral blood mononuclear cells are independent of the prostaglandin subtype. Immunology 107:152–159

    Article  CAS  Google Scholar 

  14. Hauer AC, Breese EJ, Walker-Smith JA, MacDonald TT (1997) The frequency of cells secreting interferon-gamma and interleukin-4, -5, and -10 in the blood and duodenal mucosa of children with cow’s milk hypersensitivity. Pediatr Res 42:629–638

    Article  CAS  Google Scholar 

  15. Heyman M (2005) Gut barrier dysfunction in food allergy. Eur J Gastroenterol Hepatol 17:1279–1285

    Article  Google Scholar 

  16. Hosea Blewett HJ, Cicalo MC, Holland CD, Field CJ (2008) The immunological components of human milk. Adv Food Nutr Res 54:45–80

    Google Scholar 

  17. Howie PW, Forsyth JS, Ogston SA, Clark A, Florey CD (1990) Protective effect of breast feeding against infection. BMJ 300:11–16

    Article  CAS  Google Scholar 

  18. Jacob C, Yang PC, Darmoul D, Amadesi S, Saito T, Cottrell GS, Coelho AM, Singh P, Grady EF et al (2005) Mast cell tryptase controls paracellular permeability of the intestine. Role of protease-activated receptor 2 and beta-arrestins. J Biol Chem 280:31936–31948

    Article  CAS  Google Scholar 

  19. Jiang WG, Bryce RP, Horrobin DF, Mansel RE (1998) Regulation of tight junction permeability and occludin expression by polyunsaturated fatty acids. Biochem Biophys Res Commun 244:414–420

    Article  CAS  Google Scholar 

  20. Koletzko B, Rodriguez-Palmero M (1999) Polyunsaturated fatty acids in human milk and their role in early infant development. J Mammary Gland Biol Neoplasia 4:269–284

    Article  CAS  Google Scholar 

  21. Koletzko B, Rodriguez-Palmero M, Demmelmair H, Fidler N, Jensen R, Sauerwald T (2001) Physiological aspects of human milk lipids. Early Hum Dev 65(suppl):S3–S18

    Article  CAS  Google Scholar 

  22. Li Q, Zhang Q, Wang M, Zhao S, Xu G, Li J (2008) n-3 Polyunsaturated fatty acids prevent disruption of epithelial barrier function induced by proinflammatory cytokines. Mol Immunol 45:1356–1365

    Article  CAS  Google Scholar 

  23. Li Q, Zhang Q, Zhang M, Wang C, Zhu Z, Li N, Li J (2008) Effect of n-3 polyunsaturated fatty acids on membrane microdomain localization of tight junction proteins in experimental colitis. Febs J 275:411–420

    CAS  Google Scholar 

  24. Liu AH (2001) Allergy and asthma prevention: the cup half full. Allergy Asthma Proc 22:333–336

    CAS  Google Scholar 

  25. Lopez-Pedrosa JM, Ramirez M, Torres MI, Gil A (1999) Dietary phospholipids rich in long-chain polyunsaturated fatty acids improve the repair of small intestine in previously malnourished piglets. J Nutr 129:1149–1155

    CAS  Google Scholar 

  26. Mengeaud V, Nano JL, Fournel S, Rampal P (1992) Effects of eicosapentaenoic acid, gamma-linolenic acid and prostaglandin E1 on three human colon carcinoma cell lines. Prostaglandins Leukot Essent Fatty Acids 47:313–319

    Article  CAS  Google Scholar 

  27. Mitic LL, Van Itallie CM, Anderson JM (2000) Molecular physiology and pathophysiology of tight junctions I. Tight junction structure and function: lessons from mutant animals and proteins. Am J Physiol Gastrointest Liver Physiol 279:G250–G254

    CAS  Google Scholar 

  28. Morrison WR, Smith LM (1964) Preparation of fatty acid methyl esters and dimethylacetals from lipids with boron fluoride–methanol. J Lipid Res 53:600–608

    Google Scholar 

  29. Nano JL, Nobili C, Girard-Pipau F, Rampal P (2003) Effects of fatty acids on the growth of Caco-2 cells. Prostaglandins Leukot Essent Fatty Acids 69:207–215

    Article  CAS  Google Scholar 

  30. Nusrat A, Turner JR, Madara JL (2000) Molecular physiology and pathophysiology of tight junctions. IV. Regulation of tight junctions by extracellular stimuli: nutrients, cytokines, and immune cells. Am J Physiol Gastrointest Liver Physiol 279:G851–G857

    CAS  Google Scholar 

  31. van Odijk J, Kull I, Borres MP, Brandtzaeg P, Edberg U, Hanson LA, Host A, Kuitunen M, Olsen SF et al (2003) Breastfeeding and allergic disease: a multidisciplinary review of the literature (1966–2001) on the mode of early feeding in infancy and its impact on later atopic manifestations. Allergy 58:833–843

    Article  Google Scholar 

  32. Oh IS, Shimizu H, Sato T, Uehara Y, Okada S, Mori M (2005) Molecular mechanisms associated with leptin resistance: n-3 polyunsaturated fatty acids induce alterations in the tight junction of the brain. Cell Metab 1:331–341

    Article  CAS  Google Scholar 

  33. Reynier M, Sari H, d’Anglebermes M, Kye EA, Pasero L (1991) Differences in lipid characteristics of undifferentiated and enterocytic-differentiated HT29 human colonic cells. Cancer Res 51:1270–1277

    CAS  Google Scholar 

  34. Roche HM, Terres AM, Black IB, Gibney MJ, Kelleher D (2001) Fatty acids and epithelial permeability: effect of conjugated linoleic acid in Caco-2 cells. Gut 48:797–802

    Article  CAS  Google Scholar 

  35. Roig-Perez S, Guardiola F, Moreto M, Ferrer R (2004) Lipid peroxidation induced by DHA enrichment modifies paracellular permeability in Caco-2 cells: protective role of taurine. J Lipid Res 45:1418–1428

    Article  CAS  Google Scholar 

  36. Rosella O, Sinclair A, Gibson PR (2000) Polyunsaturated fatty acids reduce non-receptor-mediated transcellular permeation of protein across a model of intestinal epithelium in vitro. J Gastroenterol Hepatol 15:626–631

    Article  CAS  Google Scholar 

  37. Rouwet EV, Heineman E, Buurman WA, ter Riet G, Ramsay G, Blanco CE (2002) Intestinal permeability and carrier-mediated monosaccharide absorption in preterm neonates during the early postnatal period. Pediatr Res 51:64–70

    Article  CAS  Google Scholar 

  38. Sanders SE, Madara JL, McGuirk DK, Gelman DS, Colgan SP (1995) Assessment of inflammatory events in epithelial permeability: a rapid screening method using fluorescein dextrans. Epithelial Cell Biol 4:25–34

    CAS  Google Scholar 

  39. Shulman RJ, Schanler RJ, Lau C, Heitkemper M, Ou CN, Smith EO (1998) Early feeding, antenatal glucocorticoids, and human milk decrease intestinal permeability in preterm infants. Pediatr Res 44:519–523

    Article  CAS  Google Scholar 

  40. Stillwell W, Wassall SR (2003) Docosahexaenoic acid: membrane properties of a unique fatty acid. Chem Phys Lipids 126:1–27

    Article  CAS  Google Scholar 

  41. Usami M, Komurasaki T, Hanada A, Kinoshita K, Ohata A (2003) Effect of gamma-linolenic acid or docosahexaenoic acid on tight junction permeability in intestinal monolayer cells and their mechanism by protein kinase C activation and/or eicosanoid formation. Nutrition 19:150–156

    Article  CAS  Google Scholar 

  42. Usami M, Muraki K, Iwamoto M, Ohata A, Matsushita E, Miki A (2001) Effect of eicosapentaenoic acid (EPA) on tight junction permeability in intestinal monolayer cells. Clin Nutr 20:351–359

    Article  CAS  Google Scholar 

  43. Vine DF, Charman SA, Gibson PR, Sinclair AJ, Porter CJ (2002) Effect of dietary fatty acids on the intestinal permeability of marker drug compounds in excised rat jejunum. J Pharm Pharmacol 54:809–819

    Article  CAS  Google Scholar 

  44. Weaver LT, Laker MF, Nelson R (1984) Intestinal permeability in the newborn. Arch Dis Child 59:236–241

    CAS  Google Scholar 

  45. Willemsen LE, Hoetjes JP, van Deventer SJ, van Tol EA (2005) Abrogation of IFN-gamma mediated epithelial barrier disruption by serine protease inhibition. Clin Exp Immunol 142:275–284

    Article  CAS  Google Scholar 

  46. Willemsen LE, Koetsier MA, van Deventer SJ, van Tol EA (2003) Short chain fatty acids stimulate epithelial mucin 2 expression through differential effects on prostaglandin E(1) and E(2) production by intestinal myofibroblasts. Gut 52:1442–1447

    Article  CAS  Google Scholar 

  47. Yamagata K, Tagami M, Takenaga F, Yamori Y, Nara Y, Itoh S (2003) Polyunsaturated fatty acids induce tight junctions to form in brain capillary endothelial cells. Neuroscience 116: 649–656

    Article  CAS  Google Scholar 

  48. Yang PC, Berin MC, Yu LC, Conrad DH, Perdue MH (2000) Enhanced intestinal transepithelial antigen transport in allergic rats is mediated by IgE and CD23 (FcepsilonRII). J Clin Invest 106:879–886

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Danone Research, Centre for Specialised Nutrition, Wageningen, The Netherlands

    Linette E. M. Willemsen PhD, Marleen A. Koetsier, Martin Balvers & Eric A. F. van Tol

  2. Pharmacology and Pathophysiology Utrecht Institute for Pharmaceutical Sciences, Utrecht University, PO Box 80082, 3508, TB, Utrecht, The Netherlands

    Linette E. M. Willemsen PhD

  3. Danone Research, Centre for Specialised Nutrition, Friedrichsdorf, Germany

    Christopher Beermann & Bernd Stahl

Authors
  1. Linette E. M. Willemsen PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marleen A. Koetsier
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Martin Balvers
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Christopher Beermann
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Bernd Stahl
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Eric A. F. van Tol
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Linette E. M. Willemsen PhD.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Willemsen, L.E.M., Koetsier, M.A., Balvers, M. et al. Polyunsaturated fatty acids support epithelial barrier integrity and reduce IL-4 mediated permeability in vitro. Eur J Nutr 47, 183–191 (2008). https://doi.org/10.1007/s00394-008-0712-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00394-008-0712-0

Keywords

Search

Navigation